IL-12α deficiency attenuates pressure overload-induced cardiac inflammation, hypertrophy, dysfunction, and heart failure progression

Figures

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  Figure 1. IL-12α KO ameliorated TAC-induced LV failure, LV hypertrophy, increase of lung weight, and RV hypertrophy in male mice. (A–D) Quantified data of echocardiographic measurements of LV ejection fraction, LV fractional shortening, LV end-systolic diameter, and LV end-diastolic diameter. (E–G) LV β-MHC and ANP expression in corresponding groups. (H–O) The ratio of LV weight, LA weight, lung weight, RV weight to tibial length or body weight in corresponding mice. *p<0.05; ns, non-significant.

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  Figure 2. IL-12α KO significantly attenuated TAC-induced LV cardiomyocyte hypertrophy, fibrosis, and leukocyte infiltration. (A, B) Representative images and quantified data of LV cardiomyocyte size determined by FITC conjugated wheat germ agglutinin (WGA) staining. (C, D) Representative images and quantified data of LV fibrosis performed by Sirius red/Fast green staining. (E–H) Representative images and quantified data of LV leukocyte or macrophage accumulation performed by immuno-histological staining. *p<0.05; ns, non-significant.

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  Figure 3. IL-12α KO significantly attenuated TAC-induced pulmonary inflammation, fibrosis, and vessel remodeling. (A–D) Representative images and quantified data of infiltrating leukocytes or macrophages in the lung performed by immuno-histological staining. (E, F) Representative images and quantified data of lung fibrosis performed by Sirius red/Fast green staining. (G, H) Representative images and quantified data of pulmonary vessel remodeling performed by immuno-histological staining. (I–L) Alterations of MCP1, IL-6, IL-1α, and IL-1β per lungs sample. *p<0.05; ns, non-significant.

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  Figure 4. IL-12α KO significantly attenuated TAC-induced lung dendritic cell (CD11chighF4/80-) accumulation and activation. (A) Flow cytometry plots for CD11chighF4/80- cells in the lung. (B) Percentage of CD11chighF4/80- cells within CD45+ leukocytes. (C) Flow cytometry plots for the identification of MHCIIhighCD11chighF4/80- dendritic cells. (D, E) Quantified percentage of MHCIIhighCD11chighF4/80- dendritic cells within CD11chighF4/80- or CD45+ leukocytes. (F) Mean fluorescent intensity of MHCII in CD11chighF4/80- cells. (G) Representative histograms showing MHCII expression in CD11chighF4/80- cells. *p<0.05, ns, non-significant.

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  Figure 5. IL-12α KO significantly attenuated TAC-induced pulmonary F4/80+ macrophage accumulation and polarization. (A) Flow cytometry plots of lung F4/80+ macrophages. (B) Quantified percentage of F4/80+ cells within CD45+ cells. (C) Flow cytometry plots for detecting MHCII expression in F4/80+ cells. (D, E) The percentages of MHCIIhighF4/80+ cells within F4/80+ or CD45+ cells. (F) Mean fluorescent intensity of MHCII in F4/80+ cells. (G) Ratio of MHCIIhigh to MHCIIlow in F4/80+ cells. (H) Representative histograms of MHCII expression in F4/80+ cells. *p<0.05, ns, non-significant.

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  Figure 6. IL-12α KO significantly ameliorated TAC-induced accumulation of lung alveolar macrophages (CD11chighCD11blowF4/80+) and their activation. (A) Flow cytometry plots for alveolar macrophages. (B) Quantified alveolar macrophages within CD45+ leukocytes. (C) Flow cytometry plots for the detection of MHCII expression in alveolar macrophages. (D–F) Quantified percentages MHCIIhigh alveolar macrophages within alveolar macrophages, F4/80+ cells, and CD45+ cells, respectively. (G) Mean fluorescent intensity of MHCII in alveolar macrophages. (H) Representative histograms showing the relative MHCII expression in each group. *p<0.05, ns, non-significant.

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  Figure 7. IL-12α KO ameliorated TAC-induced accumulation of Ly6Clow interstitial macrophages (Ly6clowCD11clowCD11bhighF4/80+) and their activation. (A) Representative gating plots of Ly6Clow interstitial macrophages. (B) Quantified percentage of Ly6Clow interstitial macrophages within CD45+ leukocytes. (C) Representative plots for MHCIIhigh macrophages within Ly6Clow interstitial macrophages. (D–F) Percentages of MHCIIhighLy6Clow interstitial macrophages within Ly6Clowinterstitial macrophages, F4/80+, and CD45+ cells, respectively. (G) Mean fluorescent intensity of MHCII in Ly6Clow interstitial macrophages. (H) Representative histogram showing MHCII expression in Ly6Clow interstitial macrophages. *p<0.05, ns, non-significant.

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  Figure 8. IL-12α KO ameliorated TAC-induced activation of lung Ly6Chigh interstitial macrophages (Ly6ChighCD11clowCD11bhighF4/80+). (A) Representative gating plots for Ly6Chigh interstitial macrophages. (B) Percentages of Ly6Chigh interstitial macrophages within CD45+ cells. (C) Representative plots for MHCIIhigh macrophages within Ly6Chigh interstitial macrophages. (D–F) Percentage of MHCIIhighLy6Chigh interstitial macrophages within the Ly6Chigh interstitial macrophages, F4/80+, and CD45+ cells, respectively. (G) Mean fluorescent intensity of MHCII in Ly6Chigh interstitial macrophages. (H) Representative histogram showing MHCII expression Ly6Chigh interstitial macrophages. *p<0.05, ns, non-significant.

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  Figure 9. IL-12α KO significantly attenuated TAC-induced T cell activation in the lung. (A) Representative plots detecting activation status of CD3+ T cells. (B–D) Percentages of CD44+CD62L- effector memory cells, CD44+CD62L+ central memory cells, and CD44-CD62L+ naïve cells within CD3+ T cells, respectively. (E) Quantified mean fluorescent intensity of CD44 in CD3+ T cells. (F) Representative histograms of CD44 expression within CD3+ T cells. (G) Representative plots detecting activation status of CD4+ T cells. (H–J) The percentages of CD44+CD62L- effector memory cells, CD44+CD62L+ central memory cells, and CD44-CD62L+ naïve cells within CD4+ T cells, respectively. (K) Mean fluorescent intensity of CD44 in CD4+ T cells. (L) Representative histograms of CD44 expression within CD4+ T cells. (M) Representative plots detecting activation status of CD8+ T cells. (N–P) The percentage of CD44+CD62L- effector memory cells, CD44+CD62L+ central memory cells, and CD44-CD62L+ naïve cells within CD8+ T cells, respectively. (Q) Mean fluorescent intensity of CD44 in CD8+ T cells. (R) Representative histograms showing CD44 expression within CD8+ T cells. *p<0.05, ns, non-significant.